Porphyromonas gingivalis outer membrane vesicles modulate cytokine and chemokine production by gingipain-dependent mechanisms in human macrophages.

OBJECTIVE: The aim was to determine the changes of inflammatory mediator expression in human macrophages stimulated with outer membrane vesicles purified from Porphyromonas gingivalis. DESIGN: outer membrane vesicles purified by ultracentrifugation from ATCC 33277 and W83 P. gingivalis strains were used for stimulating human macrophages and determine their inflammatory mediator expression changes. U937 monocyte cells line were differentiated into macrophages and stimulated with outer membrane vesicles for 30 min and six hours. In Independent experiments, the outer membrane vesicles and viable bacteria control were pre-treated with the gingipain inhibitors KYT-1 and KYT-36 (Arg-gingipain and Lys-gingipain, respectively) or Polymyxin-B to block the lipopolysaccharide activity to evaluate the secretion changes of immune mediators IL-1ß, IL-6, TNF-α, IL-8, MCP-1, MIP-1α and RANTES by flow cytometry. A factorial ANOVA was used to analyze the data. RESULTS: The outer membrane vesicles of P. gingivalis ATCC 33277 displayed higher Arg-gingipain activity than those obtained from the P. gingivalis W83 strain (0.6 U/µg vs. 0.46 U/µg). Although the outer membrane vesicles of P. gingivalis stimulated the production of cytokines and chemokines, specific Arg-gingipain and Lys-gingipain inhibition induced significant increases in IL-1ß, IL-6, IL-8, MCP-1, and RANTES levels, and this induction was significantly greater at 6 h compared to 30 min (*p < 0.05). On the contrary, TNF-α secretion decreased when gingipains were blocked. CONCLUSIONS: outer membrane vesicles may play a dual role during P. gingivalis infection based on their ability to induce changes in the immune responses of human macrophages, probably via gingipain-dependent events.

Human papilloma virus genotypes in dysplasia and epithelial hyperplasia of oral cavity using the luminex xmap technology. A multicenter study.

BACKGROUND: Oral cancer associated with high risk (HPV-HR) human papilloma virus (HPV) has been increasing. HPV-HR has been associated with epithelial dysplasia, however, little information exists on its frequency in epithelial hyperplasia lesions. The aim of this study is to compare HPV genotypes in dysplastic and hyperplastic lesions of oral cavity. MATERIAL AND METHODS: Two hundred and fifty oral lesions: 131 dysplasia and 119 hyperplasia from two regions of Colombia were evaluated. One hundred seventy-four coming from urban area and 104 from a high risk population to oral cancer from a rural area. HPV was identified by qPCR and Twenty-four HPVs genotypes were evaluated by Luminex® technology. Logistic regressions were performed to establish the associations between HPV infections with oral dysplasia. RESULTS: Twenty-eight percent (70/250) of the samples were positives for any HPV and HPV-HRs were more frequently than low risk HPVs. HPV-16 was the most detected genotype (16%) followed by HPV-31, 53, 18 and 45. HPV, HPV-HRs and HPV-16 were only associated with dysplasia in urban area; OR 3.28 (CI 95% 1.49-7.17), OR 7.94 (CI 95% 2.97-21.2) and OR 5.90 (CI 95% 2.05-17). Individuals in rural area showed more HPV and HPV-HRs infection in hyperplasic lesions than urban population. The majority of HPV+ lesions had multi-type of HPV (52/70) and the urban individuals showed more genotypes than rural population. CONCLUSIONS: HPV-.HRs are frequently found in hyperplastic and dysplastic epithelial lesions. HPV-HRs and HPV-16 were associated with dysplasia in urban population. Rural high risk population and urban population differ in the frequency and variety of HPV genotypes.

Electrostimulation of the lingual nerve by an intraoral device may lead to salivary gland regeneration: A case series study.

BACKGROUND: Salivary gland function is controlled by the salivary reflex, whose efferent arm is composed by the parasympathetic and the sympathetic divisions of the autonomic nervous system. Parenchymal injury is the main salivary gland involvement of Sjögren's syndrome and head and neck radiotherapy, but neural damage has been reported as well. Recently an intraoral device for electrostimulation of the lingual nerve in vicinity to the lower third molar has been introduced. At this point this nerve carries efferent fibers for the innervation of the submandibular, sublingual and several minor salivary glands and afferent fibers of the salivary reflex. Therefore, excitation of these fibers potentially leads to increased secretion of all salivary glands. Thus, the study objective was to assess whether comprehensive neural activation by electrostimulation of the lingual nerve carries the potential to induce the regeneration of damaged salivary glands. MATERIAL AND METHODS: The device was tested on three patients with no collectable resting and stimulated secretion of saliva during a double blind, sham controlled period of two months and nine open-label months. RESULTS: All three subjects developed the capacity to spit saliva, not only in direct response to the electrostimulation but also after free intervals without electrostimulation. In addition, their symptoms of dry mouth severity and frequency improved. CONCLUSIONS: This recovery is probably due to the combined effect of increase in secretory functional gland mass and regain of nervous control of the secretory elements and blood vessels. Both are phenomena that would contribute to gland regeneration.

Evaluation of substantivity of hypochlorous acid as an antiplaque agent: A randomized controlled trial.

BACKGROUND: Hypochlorous acid (HOCl) is a non-antibiotic antimicrobial substance with significant effects on pathogenic oral micro-organisms. The effects of HOCl as an antiplaque agent have not been studied. OBJECTIVE: The aim of this study was to evaluate the substantivity of HOCl mouthwashes compared with chlorhexidine (CHX) rinses and a placebo. MATERIALS AND METHODS: A double-blind randomized controlled trial with 75 participants was conducted. Participants were divided into five groups using block randomization: 0.025% HOCl, 0.05% HOCl, 0.12% CHX, 0.2% CHX, and sterile water as a placebo. Participants were instructed to use each rinse solution for 30 seconds after dental prophylaxis. Samples of saliva were taken at baseline and after 30 seconds, 1, 3, 5 and 7 hours to assess substantivity, and bacterial viability was established by the fluorescence method. Visible plaque in all participants was assessed with the Turesky index at baseline and at 7 hours, and adverse events were also assessed. RESULTS: HOCl led to a 33% reduction in bacterial counts in the saliva after 30 seconds compared with a 58% reduction by CHX. HOCl has no substantivity, and bacterial counts returned to baseline after 1 hour. Placebo treatment led to the highest plaque count after 7 hours compared with the CHX and HOCl groups, although the differences were not significant. HOCl rinsing induced the highest percentages of unpleasant taste and dryness sensations. CONCLUSIONS: HOCl rinses have an initial effect on bacterial viability in saliva but have no substantivity. Other mechanisms may explain its antiplaque effect.

Two-dimensional and three-dimensional models for studying atherosclerosis pathogenesis induced by periodontopathogenic microorganisms.

Epidemiological studies have established a clinical association between periodontal disease and atherosclerosis. Bacteremia and endotoxemia episodes in patients with periodontitis appear to link these two diseases by inducing a body-wide production of cardiovascular markers. The presence of oral bacteria in atherosclerotic lesions in patients with periodontitis suggests that bacteria, or their antigenic components, induce alterations in the endothelium associated with atherosclerosis. Therefore, a causal mechanism explaining the association between both diseases can be constructed using in vitro models. This review presents current experimental approaches based on in vitro cell models used to shed light on the mechanism by which periodontal pathogenic microorganisms, and their antigenic components, induce proatherosclerotic endothelial activity. Monolayer cultures of endothelial vascular or arterial cells have been used to assess periodontal pathogenic bacteria and their antigenic compounds and endothelial activation. However, these models are not capable of reflecting the physiological characteristics of the endothelium inside vascularized tissue. Therefore, the shift from two-dimensional (2D) cellular models toward three-dimensional (3D) models of endothelial cells resembling an environment close to the physiological environment of the endothelial cell within the endothelium is useful for evaluating the physiological relevance of results regarding the endothelial dysfunction induced by periodontopathogens that are currently obtained from 2D models. The use of in vitro 3D cellular models can also be relevant to the search for therapeutic agents for chronic inflammatory diseases such as atherosclerosis. Here, we present some strategies for the assembly of 3D cultures with endothelial cells, which is useful for the study of periodontopathogen-mediated disease.

Accuracy of cotinine serum test to detect the smoking habit and its association with periodontal disease in a multicenter study.

BACKGROUND: The validity of the surveys on self-reported smoking status is often questioned because smokers underestimate cigarette use and deny the habit. It has been suggested that self-report should be accompanied by cotinine test. This report evaluates the usefulness of serum cotinine test to assess the association between smoking and periodontal status in a study with a large sample population to be used in studies with other serum markers in epidemiologic and periodontal medicine researches. MATERIAL AND METHODS: 578 patients who were part of a multicenter study on blood biomarkers were evaluated about smoking and its relation to periodontal disease. Severity of periodontal disease was determinate using clinical attachment loss (CAL). Smoking was assessed by a questionnaire and a blood sample drawn for serum cotinine determination. RESULTS: The optimal cut-off point for serum cotinine was 10 ng/ml. Serum cotinine showed greater association with severity of CAL than self-report for mild-moderate CAL [OR 2.03 (CI95% 1.16-3.53) vs. OR 1.08 (CI95% 0.62-1.87) ] advanced periodontitis [OR 2.36 (CI95% 1.30- 4.31) vs. OR 2.06 (CI95% 0.97-4.38) ] and extension of CAL > 3 mm [ OR 1.78 (CI95% 1.16-1.71) vs. 1.37 (CI95% 0.89-2.11)]. When the two tests were evaluated together were not shown to be better than serum cotinine test. CONCLUSIONS: Self-reported smoking and serum cotinine test ≥ 10ng/ml are accurate ,complementary and more reliable methods to assess the patient's smoking status and could be used in studies evaluating serum samples in large population and multicenter studies. CLINICAL RELEVANCE: The serum cotinine level is more reliable to make associations with the patient's periodontal status than self-report questionnaire and could be used in multicenter and periodontal medicine studies.

Is the Treatment with Biological or Non-biological DMARDS a Modifier of Periodontal Condition in Patients with Rheumatoid Arthritis?

BACKGROUND AND OBJECTIVE: Experimental models suggest the use of different therapy protocols in rheumatoid arthritis (RA) as modulators on periodontal condition. This study evaluated the effects of conventional drug treatment and anti-TNF therapy in patients with RA on microbiological and periodontal condition, establishing the association of markers of periodontal infection with indexes of rheumatic activity. MATERIALS AND METHODS: One hundred seventy nine individuals with RA were evaluated (62 with anti-TNF-. and 115 with only DMARDs). The periodontal evaluation included plaque and gingival indexes, bleeding on probing (BOP), clinical attachment loss (CAL), pocket depth (PD) and subgingival plaque samples for microbiological analysis. Rheumatologic evaluations included a clinical examination, rheumatoid factor (RF), antibodies against cyclic-citrullinated peptides (ACPAs), and activity markers (DAS28-ERS), high sensitive C-reactive protein (hs-CRP), erythrocyte sedimentation rate (ESR). RESULTS: Anti-TNF-alpha therapy influenced periodontal microbiota with a higher frequency of T. denticola (p=0.01). Methotrexate combined with leflunomide exhibited a higher extension of CAL (p=0.005), and anti-TNF-alpha therapy with methotrexate was associated with a lower extension of CAL (p=0.05). The use of corticosteroids exerted a protective effect on the number of teeth (p=0.027). The type of DMARD affected P. gingivalis, T. forsythia and E. nodatum presence. Elevated ACPAs titers were associated with the presence of red complex periodontal pathogens (p=0.025). Bleeding on probing was associated with elevated CPR levels (p=0.05), and ESR was associated with a greater PD (p=0.044) and presence of red complex (p=0.030). CONCLUSION: Different pharmacological treatments for RA affect the clinical condition and subgingival microbiota.

An "all 5 mm ports" technique for laparoscopic day-case anti-reflux surgery: A consecutive case series of 205 patients.

INTRODUCTION: Laparoscopic anti-reflux surgery is conventionally performed using two 10/12 mm ports. While laparoscopic procedures reduce post-operative pain, the use of larger ports invariably increases discomfort and affects cosmesis. We describe a new all 5 mm ports technique for laparoscopic anti-reflux surgery and present a review of our initial experience with this approach. METHODS: All patients undergoing laparoscopic fundoplication over a 35 month period from February 2013 under the care of a single surgeon were included. A Lind laparoscopic fundoplication was performed using an all 5 mm port technique. Data was recorded prospectively on patient demographics, operating surgeon, surgical time, date of discharge, readmissions, complications, need for re-intervention, and reasons for admission. RESULTS: Two hundred and five consecutive patients underwent laparoscopic fundoplication over the study period. The all 5 mm port technique was used in all cases, with conversion to a 12 mm port only once (0.49%). Median operating time was 52 min 185 (90.2%) patients were discharged as day cases. Increasing ASA grade and the presence of a hiatus hernia were associated with the need for overnight stay with admission required in 33% of patients with ASA 3, compared to 4% with ASA 1 (p = 0.001), and 29% of those with a hiatus hernia vs. 5% without (p < 0.001). No port-related complications occurred, and no patients developed recurrence of reflux symptoms. A single patient required mesh repair of a large hiatus hernia. CONCLUSION: The all 5 mm ports approach to laparoscopic anti-reflux surgery is a safe, efficient, and cost-effective technique which facilitates same day discharge and minimises port related complications. National commissioning guidelines in the UK should target quality improvements in anti-reflux surgery based around day-case management. This would improve the service for these patients and culminate in cost savings for the NHS.

fimA genotypes and PFGE profile patterns in Porphyromonas gingivalis isolates from subjects with periodontitis.

BACKGROUND AND OBJECTIVES: Porphyromonas gingivalis is frequently identified to type by evaluation of fimA polymorphisms and less often by pulsed-field gel electrophoresis (PFGE) because of the technical intricacies of PFGE. To compare these techniques, we genotyped P. gingivalis clinical isolates as to (i) their fimA type and (ii) their whole genome restriction profile (PFGE analysis). MATERIAL AND METHODS: Thirty-two P. gingivalis strains were isolated from 16 unrelated periodontitis patients. Two strains were isolated from each patient. Strains were subjected to a fimA-typing polymerase chain reaction (PCR) assay. Strains that could not be typed by PCR were submitted to sequencing of the entire fimA gene. The PFGE profiles of clinical strains were compared using bioinformatic analysis. RESULTS: Seven of the 32 isolates were not typeable by PCR and so their entire fimA gene was sequenced. The sequencing identified each strain as belonging to a single fimA type. In one case, sequencing of the fimA gene did not agree with the result obtained using fimA PCR typing. With the exception of one patient, each patient presented isolates bearing the same fimA type. However, in three patients, isolates with the same fimA type presented different PFGE pulsotypes. CONCLUSION: The P. gingivalis typing using fimA PCR has limitations in typeability and discriminatory power. A typing technique for P. gingivalis that is easy to perform but that presents adequate typeability and discriminatory power is needed if we want to better understand the epidemiology of periodontal disease.

Detección de porphyromonas gingivalis y bacilos entéricos en pacientes con periodontitis crónica severa en una muestra de población de Bogotá / Detection of porphyromonas gingivalis and enteric bacilli in patients with severe chronic periodontitis in a sample of population of Bogota City

El propósito de este estudio fue evaluar la frecuencia de P. Gingivales en placa subgingival en una muestra de pacientes colombianos, residentes en Bogotá, con diagnóstico confirmado de periodontitis crónica severa, ppr medio de la Reacciión en Cadena de Polimerasa (PCR) y el aislamiento de bacilos entéricos por cultivo.

Evaluación, de la estabilidad dimensional de injertos de tejido conectivo. Un estudio a 50 días / Evaluation of the dimensional stability of connective tissue grafts. A study of 150 days

El propósito de este estudio fue evaluar la estabilidad dimensional de los injertos de tejido conectivo utilizados en la técnica de aumento de reborde con defecto tipo B de acuerdo a la clasificación de Allen y col en 1985. Participaron un total de ocho pacientes adultos, con un rango de edad entre 19 y 50 años, siete de sexo masculino y uno de sexo femenino, quienes recibieron injertos de tejido conectivo con grosores que variaron entre 1.5 mm y 3 mm siguiendo la técnica descrita por Langer y Calagna en 1980. Para evaluar la estabilidad dimensional se utilizó una férula con una guía de inserción cilíndrica que permitió la entrada de una sonda de 0.5 mm de diámetro ubicada en el área de mayor depresión. Las medidas fueron trasladadas a un Nonium que transcribió las medidas a centésimas de milímetro. Para la toma de medidas se realizó una calibración Inter. e intraexaminador y se escogió al examinador más calibrado para tomar todas las medidas del estudio, las cuales fueron realizadas en pacientes y modelo de estudio tomado de impresiones que fueron estandarizadas de acuerdo al tipo de alginato y proporciones polvo-líquido según la casa fabricante. Las medidas fueron tomadas a nivel prequirúrgico a los 8, 15, 30, 45, 60, 75, 90, 105, 120, 135 y 150 días. Los resultados demostraron que la variabilidad en la estabilidad dimensional pareció estar relacionada con el grosor de los injertos ya que la contracción de los injertos menores de 2 mm varió entre 37 y 73 y la contracción de los injertos mayores a 2 mm varió entre 9.5 y 28. Los injertos de menor grosor mostraron una contracción primaria más uniforme entre los primeros 45 días con una contracción secundaria leve y progresiva, mientras los injertos de mayor grosor fueron más variables y no siguieron un patrón uniforme llegando a picos de contracción entre 60 y 90 días seguidos por una contracción secundaria leve y progresiva. Sólo dos de los ocho casos mostraron una completa estabilización entre 75 y 120 días mientras que los seis casos restantes mostraron una contracción leve y progresiva aún a los 150 días sin observarse una completa estabilización en el período de observación.

Cambio histológicas en capuchones pericoronarios de terceros molares incluidos en diferentes estadios de nolla / Histological changes in dental follicles of impacted thrid molars in different Nolla stages

Desde hace varios años se han escrito generalidades sobre el folículo dental encontrándose descripciones como la que el saco dental es un término reservado para el tejido que separa el órgano del esmalte en desarrollo y más tarde a la corona. Ten cate (1) y genneser (2) 1985 describieron el saco dental como mesénquima circundante al órgano dental. El objetivo de este estudio fue el de realizar una descripción histológica de los cambios celulares en folículos dentales de terceros molares incluidos en diferentes estadios de nolla, entendiéndose estadio de nolla como los grados de calificación que se presentan durante el período de formación dental.

Aspectos microbiológicos y población celular en mujeres puberales y post-puberales con diagnóstico de gingivitis / Microbiologic aspects and cell population in puberty and pre puberty women with gingivitis diagnosis

El propósito este fue comprar la población celular a nivel de tejido conectivo, los morfotipos bacteriales en placa supragingival y la presencia de prevotella intermedius en placa subgingival en niñas sanas y con diagnóstico de gingivitis en etapa puberal y post-puberal. Se estudiaron 200 pacientes de los cuales se seleccionaron 85; 43 en etapa puberal. Se tomaron muestras de placas supragingival, placa subgingival y biopsiasde tejido conectivo gingival a cada paciente. Se observo en placa supragingival aumento bacilos gra(-) en pacientes con gingivitis gramo III en etapa puberal, siendo estadísticamente significativo. En placa subgingival se evidenció la presencia de prevotella intermedius solo en pacientes en con gingivitis grado III de severidad, existiendo diferencias estadísticamente significativas entre el grupo puberal y post-puberal. Se encontró una relación linfocitos: Plasmocitos de 1:5 sin encontrarse diferencias estadísticamente significativas entre los dos grupos, observándose una pérdida de correlación y la población celular.

Estudios descriptivos de morbilidad oral y antecedentes odontológicos en pacientes infectados por virus de inmunodeficiencia humana VIH, en estadios II, III y IV pertenencias al programa de atención al Sida del hospital regional Simón Bolívar en el período comprendido entre febrero y mayo de 1992 / Descriptive studies of oral morbility and dental antecedents in patients infected by Human Immunodeficiency Virus HIV, in II, III and IV stages pertaining to the Program of attention to AIDS of the Simon Bolivar Hospital, from Febrary to May, 1992

El propósito del presente estudio fue describir las alteraciones orales encontradas en pacientes VIH+ en diferentes estadios de acuerdo a las clasificaciones de OMS. Se pretendió también identificar los antecedentes odontológicos los últimos seis meses de estos pacientes. De 514 pacientes que habían asistido al programa de atención al sida del hospital Simón Bolivar hasta mayo de 1992, se avaluaron 56 en diferentes estudios. En estudios tempranos la única alteración encontrada en algunos de los pacientes fueron las linfoadenopatías. Las alteraciones en tejidos blandos fueron aparentes es estado IV, siendo las candidiasis oral la patología más observada, seguida por la leoucoplasia vellosa.El herpes simple, úlceras inespecíficas, el eritema difuso de paladar blando y el sarcoma de keposi fueron observadas en menor proporción .De 56 pacientes, 44 fueron interrogados sobre sus antecedentes odontológicos y todos los pacientes habían asistido a consulta odontológicas en los últimos 6 meses. El 40 conocía su diagnóstico y lo reportaron al odontólogo. El 20 conocía su diagnóstico y el 10 lo informaron y el 38 no conocía su diagnóstico cuando asistieron a la consulta. El 86 de los odontólogos que trataron l a los pacientes utilizaron medidas completas de bioseguridad y el 14 tomaron medidas incompletas o no las tomaron, observándose un gran impacto de la prevención al Sida en el medio odontológico.